The central objective of the Proteomics Core is to provide proteomics expertise and state-of-the-art resources for proteomics data acquisition, mining, and interpretation available through the National Center for Proteomics Research for Projects l-lll to test individual hypotheses. Each individual project will utilize the resources of the Proteomics Core in their specific aims, and the proteomic profiling studies are critical components of each individual Project. The Proteomics Core will interact with all projects to assist in sample preparation, quality control before and during proteomic experiments, and with the analysis and interpretation of data. The established infrastructure of the National Center for Proteomics Research at MCW has developed optimized tools ideally suited for the quantitative analysis of proteomes and the characterization of changes in protein phosphorylation. These tools, resources, and expertise will advance all three projects effectively and generate unique datasets highlighting the complex underlying mechanisms of anesthetic preconditioning. For this PPG, the Proteomics Core will 1) Establish and improve standard analytical procedures for mass spectrometric analysis and quantification of cellular sub-proteomes and protein phosphorylation. 2) Perform mass spectrometric analyses of mitochondrial and cytosolic samples provided by the investigators of Projects l-lll. 3) Quantify proteomic changes using isotopic labeling and tandem mass spectrometry. 4) Analyze changes in protein phosphorylation using mass spectrometric approaches. 5) Provide reliable, reproducible, uniform and timely analyses of proteomic samples for all three projects. 6) Maintain databases and automated analysis procedures for protein identification, quantification, and pathway analysis. 7) Provide expertise and equipment to assist investigators in data analysis and interpretation.